US3715188A - Cholesterol assay and reagents therefor - Google Patents
Cholesterol assay and reagents therefor Download PDFInfo
- Publication number
- US3715188A US3715188A US00139266A US3715188DA US3715188A US 3715188 A US3715188 A US 3715188A US 00139266 A US00139266 A US 00139266A US 3715188D A US3715188D A US 3715188DA US 3715188 A US3715188 A US 3715188A
- Authority
- US
- United States
- Prior art keywords
- cholesterol
- chromophore
- sample
- alcohol
- formamide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 239000003153 chemical reaction reagent Substances 0.000 title claims description 32
- 238000008620 Cholesterol Assay Methods 0.000 title abstract description 9
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 197
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 92
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims abstract description 58
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 claims abstract description 46
- 238000005259 measurement Methods 0.000 claims abstract description 29
- 239000000126 substance Substances 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 24
- 238000012360 testing method Methods 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims description 62
- 230000008569 process Effects 0.000 claims description 21
- 210000002966 serum Anatomy 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 16
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 14
- WFDIJRYMOXRFFG-UHFFFAOYSA-N acetic acid anhydride Natural products CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 9
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 claims description 8
- 239000012620 biological material Substances 0.000 claims description 7
- 230000006872 improvement Effects 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- XFRVVPUIAFSTFO-UHFFFAOYSA-N 1-Tridecanol Chemical compound CCCCCCCCCCCCCO XFRVVPUIAFSTFO-UHFFFAOYSA-N 0.000 claims description 5
- 229940087291 tridecyl alcohol Drugs 0.000 claims description 5
- 125000004432 carbon atom Chemical group C* 0.000 claims description 3
- 238000002798 spectrophotometry method Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 abstract description 9
- 230000000903 blocking effect Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 11
- 238000003556 assay Methods 0.000 description 9
- 239000002253 acid Substances 0.000 description 8
- 238000011161 development Methods 0.000 description 7
- 229960000583 acetic acid Drugs 0.000 description 5
- UVYVLBIGDKGWPX-KUAJCENISA-N digitonin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)C[C@@H](O)[C@H](O[C@H]5[C@@H]([C@@H](O)[C@@H](O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)CO7)O)[C@H](O)[C@@H](CO)O6)O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O7)O)[C@@H](O)[C@@H](CO)O6)O)[C@@H](CO)O5)O)C[C@@H]4CC[C@H]3[C@@H]2[C@@H]1O)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 UVYVLBIGDKGWPX-KUAJCENISA-N 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 230000002452 interceptive effect Effects 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- QRLVDLBMBULFAL-UHFFFAOYSA-N Digitonin Natural products CC1CCC2(OC1)OC3C(O)C4C5CCC6CC(OC7OC(CO)C(OC8OC(CO)C(O)C(OC9OCC(O)C(O)C9OC%10OC(CO)C(O)C(OC%11OC(CO)C(O)C(O)C%11O)C%10O)C8O)C(O)C7O)C(O)CC6(C)C5CCC4(C)C3C2C QRLVDLBMBULFAL-UHFFFAOYSA-N 0.000 description 4
- 150000001298 alcohols Chemical class 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- UVYVLBIGDKGWPX-UHFFFAOYSA-N digitonine Natural products CC1C(C2(CCC3C4(C)CC(O)C(OC5C(C(O)C(OC6C(C(OC7C(C(O)C(O)CO7)O)C(O)C(CO)O6)OC6C(C(OC7C(C(O)C(O)C(CO)O7)O)C(O)C(CO)O6)O)C(CO)O5)O)CC4CCC3C2C2O)C)C2OC11CCC(C)CO1 UVYVLBIGDKGWPX-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 238000012937 correction Methods 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 235000005911 diet Nutrition 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000007127 saponification reaction Methods 0.000 description 3
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 description 2
- 208000024799 Thyroid disease Diseases 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- MWKFXSUHUHTGQN-UHFFFAOYSA-N decan-1-ol Chemical compound CCCCCCCCCCO MWKFXSUHUHTGQN-UHFFFAOYSA-N 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000000873 masking effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 125000002889 tridecyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- UEWGGKWOHRJILY-KPNWGBFJSA-N (3S,8S,9S,10R,13R,14S,17R)-10,13-dimethyl-17-[(2R)-6-methylheptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-ol formamide Chemical compound [C@@H]1(CC[C@H]2[C@@H]3CC=C4C[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C)[C@H](C)CCCC(C)C.C(=O)N UEWGGKWOHRJILY-KPNWGBFJSA-N 0.000 description 1
- RLHIRZFWJBOHHD-UHFFFAOYSA-N 3,5-cholestadiene Natural products C1C=C2C=CCCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 RLHIRZFWJBOHHD-UHFFFAOYSA-N 0.000 description 1
- QDTDKYHPHANITQ-UHFFFAOYSA-N 7-methyloctan-1-ol Chemical compound CC(C)CCCCCCO QDTDKYHPHANITQ-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- RLHIRZFWJBOHHD-HKQCOZBKSA-N Cholesta-3,5-diene Chemical compound C1C=C2C=CCC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 RLHIRZFWJBOHHD-HKQCOZBKSA-N 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- 206010020961 Hypocholesterolaemia Diseases 0.000 description 1
- 239000004439 Isononyl alcohol Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- TUCNEACPLKLKNU-UHFFFAOYSA-N acetyl Chemical compound C[C]=O TUCNEACPLKLKNU-UHFFFAOYSA-N 0.000 description 1
- DGOBMKYRQHEFGQ-UHFFFAOYSA-L acid green 5 Chemical compound [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 DGOBMKYRQHEFGQ-UHFFFAOYSA-L 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- IBDXZWQCLMSDKQ-FDXOKOSPSA-N i-cholesterol Chemical compound C([C@H]1[C@@H]2CC[C@@H]([C@]2(CC[C@@H]1[C@@]1(C)CC2)C)[C@H](C)CCCC(C)C)[C@@H](O)[C@@]31[C@H]2C3 IBDXZWQCLMSDKQ-FDXOKOSPSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000007430 reference method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 125000002328 sterol group Chemical group 0.000 description 1
- 150000003460 sulfonic acids Chemical class 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 208000021510 thyroid gland disease Diseases 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
- Y10T436/25375—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
- Y10T436/255—Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.] including use of a solid sorbent, semipermeable membrane, or liquid extraction
Definitions
- ABSTRACT A cholesterol assay, using formamide, which achieves the simplicity of a direct test of a specimen sample, yet which also achieves a comparable specificity and accuracy of cholesterol assays involving multiple-extraction steps.
- the formamide destroys the cholesterol chromophore of a blank measurement, and this result may be compared with a companion cholesterol assay to eliminate the error caused by the chromophore of non-cholesterol chromogenic substances of the specimen; and the formamide also keeps the alcohol, which is used to prevent turbidity, from blocking the formation of the bilirubin portion of the noncholesterol chromogenic substances.
- Cholesterol which is one of the fatty materials found in the blood serum, and which is formed primarily in the liver, is generally acknowledged to be very significant as a factor in various disease processes of the human body, including diseases of the heart, vascular system, etc.
- the serum cholesterol level of an individual important but also it is important that the accuracy of the assay be close enough so as to reliably so detect any change which might occur from day to day, week to week, or month to month; since it is important to follow drug and diet therapy, and to detect cholesterol changes induced thereby, and/or changes of body response. If the test performed is not sufficiently reliable and specific, variation in some interfer ing substance might be falsely interpreted as a trend in cholesterol level, particularly since the interfering substances themselves change in concentration from time to time, and not at all always proportionately to any change of cholesterol.
- Cholesterol reacts with strongly acid reagents to produce colored substances, chiefly cholestadiene sulfonic acids.
- acetic acid and acetic anhydride are used as solvents and dehydrating reagents
- sulfuric acid or sulfuric acid and ptoluenesulfonic acids are used as dehydrating and ox idizing reagents.
- cholesterol is first attacked by strongly acid reagents, generalized as HX, where X" would be, for example, the sulfate ion or the acetyl radical.
- HX strongly acid reagents
- Such reagents first remove a molecule of water, then oxidize the intermediate to produce 3,5- cholestadiene (two double bonds).
- the oxidizing agent is usually sulfuric acid, which is converted to sulfur dioxide.
- the cholestadiene is attacked still further to form the dimer, bis-cholesta-3, S-diene, and is finally converted by the excess sulfuric acid, to a monoor disulfonic acid which is a highly colored molecule.
- TWO-STEP PROCEDURES These methods introduce an extraction step, which extracts and partially isolates the cholesterol chromogen before chromophore or color development. For this reason, some of the interfering chromogens, especially protein, are removed, but the methods still are subject to error caused by non-specific chromogens, such as bilirubin which is extracted along with the cholesterol, and thus still provides a chromophore which is mistakenly measured as cholesterol chromophore.
- Carr and Drekter that of Carr and Drekter"(7. Carr, 1.1., and Drekter, 1.1.: Clin Chem. 2:353, 1956.) is perhaps the best. Except for highly icteric samples (high bilirubin levels), it gives results that are in close correspondence to those obtained by more precise three-step methods; however, icteric samples are encountered with enough frequency as to create problems in routine analysis.
- THREE-STEP PROCEDURES These involve an extraction of the cholesterol followed by a saponification of the esters before color development. Consequently, they do not suffer from serious error because of protein. Furthermore, the saponification step tends to destroy nonspecific chromogens such as bilirubin, resulting in significantly increased accuracy over two-step methods. This class of procedures is best exemplified by the method of Abell et al.;*- (8.
- FOUR-STEP PROCEDURES These are the most complicated, but the most reliable procedures of the prior art, if their complexity does not introduce errors-which that very complexity makes possible.
- the cholesterol is extracted, the esters are saponified, and the total cholesterol is then further purified by collection as the digitonide.
- the digitonide is decomposed by saponification, which again frees the cholesterol, and the product of this step is subject to color development.
- the present invention achieves a solution to goals which heretofore have appeared to be in effect contradictory to one another; that is, as now discussed, the present invention achieves substantially the simplicity of a direct test while nevertheless achieving specificity and accuracy comparable to those involving multipleextraction steps.
- Formamide when present in a cholesterol reagent destroys the chromophore-forming reaction of the cholesterol chromogen at room temperature; however, the bilirubin chromogen does not completely form its chromophore as required for proper blank correction, at room temperature. It has been found that the formamide-cholesterol reagent must reach an elevated'temperature of about 80 C before sufficient chromophore formation from the bilirubin chromogen reaction will take place.
- Formamide and a high molecular weight alcohol thus serve the desired function of preventing cholesterol chromophore development while allowing bilirubin chromophore development in a way in which turbidity is not observed; however, since the high molecular weight alcohols do not mix with the formamide, the formamide and alcohol must be added in two separate steps to the blank. This separateness of procedural steps is avoided in the present invention by the addition of the high molecular weight alcohol to the glacial acetic acid acetic acid sulfuric acid reagent.
- the reagents are used in the following assay using a blank tube and a test tube:
- the absorbence of the test so measured is proportional to the concentration of cholesterol in serum, for the interfering chromogens contribute equally to both test and blank; and since the blank is set at zero absorhence, the absorbence of the test is a measurement of the difference between (or in effect a subtraction of) blank from test.
- 5TH EMBODIMENT Blank Reagent l Octyl, .nonyl, decyl, tridecyl, or hexadecyl alcohol.
- Blank Reagent ll Formamide
- Cholesterol Reagent As in lst Embodiment, except the alcohol is omitted.
- the reagents are used in the following assay using a blank tube and a test tube;
- An assay according to the novel concepts of the invention thus provides the multiple advantages of both simplicity corresponding to direct or one-step tests and accuracy of multiple-step or multiple-extraction tests, and also avoiding certain error-contributing aspects of the multiple-step processes.
- the often-incompatible goals of both speed and accuracy are provided, along with relative ease and convenience of the test procedures, elimination of time and error of extraction steps while nevertheless attaining in effect the result of having fully performed such extractions to avoid a masking or other effect of the other chromophoreforming chromogens usually present in the sample or specimen being assayed.
- the present invention provides a new and useful assay yielding quantitative determination of cholesterol in serum, plasma, or other biological material to be tested, and provides a method and reagents therefor, all having desired advantages and characteristics, and accomplishing the objects of the invention, including the objects and advantages hereinbefore pointed out and 'others which are inherent in the invention.
- a process for the quantitative colorimetric or spectrophotometric determination of cholesterol in serum, plasma, or other biological material to be tested wherein the improvement comprises adding formamide to said material for preventing the formation of a chromophore from the cholesterol chromogen in the sample being assayed.
- a process for the quantitative determination of cholesterol in serum, plasma, or other biological material to be tested wherein the improvement com- 0 prises adding formamide and a high molecular weight alcohol to said material for preventing the formation of chromophore from the cholesterol chromogen in the sample being assayed, while permitting the formation of the chromophore from the bilirubin chromogen.
- a process for the quantitative determination of cholesterol in serum, plasma, or other biological material to be tested wherein the improvement comprises adding formamide to block the formation of the chromophore from the cholesterol chromogen in a first portion of the sample while causing the non-cholesterol chromogenic substances in the said first portion of the sample to produce their chromophores,
- tridecl should read tridecyl This certificate supersedes Certificate of Correction d -October 23, 1973.
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Endocrinology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13926671A | 1971-04-30 | 1971-04-30 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3715188A true US3715188A (en) | 1973-02-06 |
Family
ID=22485846
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US00139266A Expired - Lifetime US3715188A (en) | 1971-04-30 | 1971-04-30 | Cholesterol assay and reagents therefor |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US3715188A (https=) |
| CA (1) | CA964174A (https=) |
| DE (1) | DE2221158C3 (https=) |
| FR (1) | FR2136763A5 (https=) |
| GB (1) | GB1393724A (https=) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3853473A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for urea determination |
| US3884638A (en) * | 1973-08-01 | 1975-05-20 | Damon Corp | Method of determining cholesterol |
| US3894844A (en) * | 1974-01-31 | 1975-07-15 | American Cyanamid Co | Simultaneous determination of triglycerides, cholesterol and phospholipids |
| US5380667A (en) * | 1992-10-30 | 1995-01-10 | The United States Of America As Represented By The Secretary Of The Air Force | Serum bilirubin and liver function tests as risk predictors for coronary artery disease |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3558516A (en) * | 1968-05-08 | 1971-01-26 | Dow Chemical Co | Cholesterol determination reagent of ferric perchlorate,sulfuric acid and ethyl acetate |
| US3615232A (en) * | 1970-02-05 | 1971-10-26 | Research Corp | Method and reagent for determining total cholesterol in blood serum |
-
1971
- 1971-04-30 US US00139266A patent/US3715188A/en not_active Expired - Lifetime
-
1972
- 1972-03-22 CA CA137,803A patent/CA964174A/en not_active Expired
- 1972-04-20 GB GB1829872A patent/GB1393724A/en not_active Expired
- 1972-04-27 FR FR7215138A patent/FR2136763A5/fr not_active Expired
- 1972-04-28 DE DE2221158A patent/DE2221158C3/de not_active Expired
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3558516A (en) * | 1968-05-08 | 1971-01-26 | Dow Chemical Co | Cholesterol determination reagent of ferric perchlorate,sulfuric acid and ethyl acetate |
| US3615232A (en) * | 1970-02-05 | 1971-10-26 | Research Corp | Method and reagent for determining total cholesterol in blood serum |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3853473A (en) * | 1972-08-30 | 1974-12-10 | Medico Electronic Inc | Reagent and method for urea determination |
| US3884638A (en) * | 1973-08-01 | 1975-05-20 | Damon Corp | Method of determining cholesterol |
| US3894844A (en) * | 1974-01-31 | 1975-07-15 | American Cyanamid Co | Simultaneous determination of triglycerides, cholesterol and phospholipids |
| US5380667A (en) * | 1992-10-30 | 1995-01-10 | The United States Of America As Represented By The Secretary Of The Air Force | Serum bilirubin and liver function tests as risk predictors for coronary artery disease |
Also Published As
| Publication number | Publication date |
|---|---|
| DE2221158A1 (de) | 1972-11-16 |
| DE2221158C3 (de) | 1981-11-12 |
| DE2221158B2 (de) | 1981-03-12 |
| FR2136763A5 (https=) | 1972-12-22 |
| GB1393724A (en) | 1975-05-14 |
| CA964174A (en) | 1975-03-11 |
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